Genetic Variation of Flavonols Quercetin, Myricetin, and Kaempferol in the Sri Lankan Tea (Camellia sinensis L.) and Their Health-Promoting Aspects.

Flavonol glycosides in tea leaves have been quantified as aglycones, quercetin, myricetin, and kaempferol. Occurrence of the said compounds was reported in fruits and vegetable for a long time in association with the antioxidant potential. However, data on flavonols in tea were scanty and, hence, this study aims to envisage the flavonol content in a representative pool of accessions present in the Sri Lankan tea germplasm. Significant amounts of myricetin, quercetin, and kaempferol have been detected in the beverage type tea accessions of the Sri Lankan tea germplasm. This study also revealed that tea is a good source of flavonol glycosides. The Camellia sinensis var. sinensis showed higher content of myricetin, quercetin, and total flavonols than var. assamica and ssp. lasiocalyx. Therefore flavonols and their glycosides can potentially be used in chemotaxonomic studies of tea germplasm. The nonbeverage type cultivars, especially Camellia rosaflora and Camellia japonica Red along with the exotic accessions resembling China type, could be useful in future germplasm studies because they are rich sources of flavonols, namely, quercetin and kaempferol, which are potent antioxidants. The flavonol profiles can be effectively used in choosing parents in tea breeding programmes to generate progenies with a wide range of flavonol glycosides.

New Sample Preparation Method for Quantification of Phenolic Compounds of Tea (Camellia sinensis L. Kuntze): A Polyphenol Rich Plant.

Chemical analysis of the Sri Lankan tea (Camellia sinensis, L.) germplasm would immensely contribute to the success of the tea breeding programme. However, the polyphenols, particularly catechins (flavan-3-ols), are readily prone to oxidation in the conventional method of sample preparation. Therefore, optimization of the present sample preparation methodology for the profiling of metabolites is much important. Two sample preparation methodologies were compared, fresh leaves (as in the conventional procedures) and freeze-dried leaves (a new procedure), for quantification of major metabolites by employing two cultivars, one is known to be high quality black tea and the other low quality black tea. The amounts of major metabolites such as catechins, caffeine, gallic acid, and theobromine, recorded in the new sampling procedure via freeze-dried leaves, were significantly higher than those recorded in the conventional sample preparation procedure. Additionally new method required less amount of leaf sample for analysis of major metabolites and facilitates storage of samples until analysis. The freeze-dried method would be useful for high throughput analysis of large number of samples in shorter period without chemical deterioration starting from the point of harvest until usage. Hence, this method is more suitable for metabolite profiling of tea as well as other phenol rich plants.

Quantification of gamma-aminobutyric acid in Sri Lankan tea by means of ultra performance tandem mass spectrometry.

Gamma-Aminobutyric acid (GABA), an important bioactive component of tea, acts as a major inhibitory neurotransmitter and is considered to influence other physiological processes in human as well as in planta. In the hereby presented study, the content of this valuable metabolite was investigated in two novel types of Ceylon Tea, explicitly "Silver Tips" and "White Tea", originating from minimally processed buds of the unique cultivar, "TRI 2043". The samples were subjected to hot water infusion, equivalent to the traditional beverage preparation procedure, and analyzed by means of hydrophilic interaction ultra performance liquid chromatography coupled to tandem mass spectrometry (HILIC LC-MS/MS). The registered GABA levels were compared with those obtained for the classic "Black Tea" and "Green Tea" samples from Sri Lanka. A high variation of GABA content was observed among the different tea types, especially in the case of "Silver Tips" and "White Tea", indicating the crucial influence of the manufacturing procedure (processing extent) on the final abundance of the bioactive component of interest. Furthermore, "White Tea" samples boasted the highest GABA concentration reported for this type of tea so far, reaching up to 50% of that characteristic of the high-priced "GABA Tea". Therefore, "White Tea" and "Silver Tips" were proved to be high quality tea with amounts of gamma-aminobutyric acid comparable with those described for similar types before. To our knowledge, this is the first report on HILIC LC-MS/MS application for the quantification of GABA and for in-depth characterization of teas from Sri Lanka.

Biosynthesis of flavan 3-ols by leucoanthocyanidin 4-reductases and anthocyanidin reductases in leaves of grape (Vitis vinifera L.), apple (Malus x domestica Borkh.) and other crops.

Catechin and epicatechin biosyntheses were studied of grape (Vitis vinifera L.), apple (Malus x domestica Borkh.) and other crop leaves, since these monomers and the derived proanthocyanidins are important disease resistance factors. Grape and apple leucoanthocyanidin 4-reductase (LAR; EC 1.17.1.3) enzymes were characterized on basis of plant and recombinant enzymes. In case of grape, two LAR cDNAs were cloned by assembling available EST sequences. Grape and apple leaf anthocyanidin reductase (ANR; EC 1.3.1.77) cDNAs were also obtained and the respective plant and recombinant enzymes were characterized. Despite general low substrate specificity, within the respective flavonoid biosyntheses of grape and apple leaves, both enzyme types deliver differently hydroxylated catechins and epicatechins, due to substrate availability in vivo. Furthermore, for LAR enzymes conversion of 3-deoxyleucocyanidin was shown. Beside relevance for plant protection, this restricts the number of possible reaction mechanisms of LAR. ANR enzyme activity was demonstrated for a number of other crop plants and its correlation with (-)-epicatechin and obvious competition with UDP-glycosyl:flavonoid-3-O-glycosyltransferases was considered.

Preformed and induced chemical resistance of tea leaf against Exobasidium vexans infection.

Levels of (-)-epicatechin in tea cultivars resistant to blister blight leaf disease were significantly higher than those in susceptible cultivars, while the reverse was true for (-)-epigallocatechin gallate, suggesting that epicatechin was involved in the resistance mechanism. The content of the methylxanthines, caffeine and theobromine, in the leaf increased in the initial translucent stage of the disease, probably as a defense response to fungal attack. Epicatechin and epigallocatechin levels were less than in healthy tissues at this stage, but increases in the corresponding gallate esters suggested that they were being converted into esters. Although epicatechin and epigallocatechin levels decreased from translucent to mature blister stages, the decrease was not significant. The decrease in levels of epicatechin, epigallocatechin, and their esters on infection and the formation of cyanidin and delphinidin on oxidative depolymerization of the blisters suggests that proanthocyanidins may play a role in the defense mechanism. The high resistance of a purple green leafed cultivar is attributed to the additional catechin source provided by the high levels of anthocyanins present.